Topical pharmaceutical composition based on semifluorinated alkanes

ABSTRACT

The invention provides liquid or semi-solid pharmaceutical compositions for topical administration comprising a semifluorinated alkane. The compositions are useful for the delivery of active ingredients into the deep layers of the skin or skin appendages. Various active ingredients may be incorporated, such as immunosuppressants, antiinfectives, antifungal agents, antiinflammatory agents, and retinoids.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No.14/122,044, filed on Nov. 25, 2013, which is a National Stage Entryunder 35 U.S.C. § 371 claiming benefit of PCT Application No.PCT/EP2012/059787, filed on May 24, 2012, which claims priority fromEuropean Application No. 11167732.4, filed May 26, 2011, and EuropeanApplication No. 11167552.6, filed May 25, 2011, the contents of each ofwhich are hereby incorporated by reference in their entireties.

BACKGROUND OF THE INVENTION

The present invention is in the field of pharmacotherapy. Morespecifically, it relates to the treatment of diseases and conditionsaffecting the skin and skin appendages.

The pharmacological treatment of cutaneous disorders may involvesystemic or local administration of an effective medicament. Typically,whenever local (e.g. epicutaneous) administration is possible and leadsto the desired outcome, it is preferred over systemic therapy which is,quite plausibly, more often associated with adverse systemic effects.However, effective treatment via the topical route of administrationrequires that the drug substance is soluble in, and/or otherwisecompatible with a suitable carrier or vehicle, and that it reaches itstarget tissue layer(s) in an effective amount or concentration. In fact,in some cases, effective epicutaneous treatment is particularlydifficult to achieve.

The skin consists of three main layers—the epidermis, the dermis and thehypodermis. The epidermis comprises the stratum corneum (the outermostsuperficial layer of skin, consisting of dead cells) and the vital(living) epidermis. Depending on the cutaneous disease or condition, oneor more layers of the skin may become affected. In some dermatologicalconditions, such as ichthyosis, callus, or plaque psoriasis, the stratumcorneum is thicker than in the healthy state and can provide asignificantly greater barrier to the penetration of a therapeuticalcompound, thus reducing its efficacy in the treatment of the deeperlayers of the skin. Moreover, it has been shown recently that withincreasing age, the skin of a human also becomes more resistant to thepenetration by drug substances.

The use of penetration enhancers has been proposed in order to achieve asufficient and consistent availability of various therapeutic compoundsat the site of action during the treatment of cutaneous diseases. Forexample, U.S. Pat. No. 5,326,566 describes a composition of apharmacological agent in combination with dibutyl adipate, or a mixtureof dibutyl adipate and isopropyl myristate, which could enhance thepenetration through the skin and even increase the amount absorbed intothe systemic circulation, if that is desired.

Even though a variety of penetration enhancers have been used forenhancing the absorption of therapeutic agents into and through theskin, substantial problems may arise when the penetration enhancers areincompatible with a particular drug substance, leading to druginstability and degradation into potentially harmful degradants.

US 2005/0079210 A1 proposes the use of liposomes the epicutaneousadministration of drugs and cosmetically useful agents. However,liposomes are difficult to manufacture cost-effectively and in areproducible manner.

Other novel formulation strategies for topical medications require theocclusion of the skin after administration, which is however perceivedas inconvenient by many patients

EP0670159A1 discloses stable oily emulsions comprising surfactant andsemifluorinated alkane, which are proposed to be suitable fortransporting oxygen to the skin or vascular system or for use as astandard for blood-gas analysis. The use of these compositions ascarriers of active agent was also suggested, although no mention is madeof whether these compositions are capable of effectively deliveringactive agents in particular to the deeper layers of the skin.

There clearly remains a need for pharmaceutical formulations andvehicles which allow the effective treatment of cutaneous disorders bytopical administration. It is therefore an object of the presentinvention to provide such improved compositions which overcome one ormore disadvantages of known compositions. In particular, it is an objectof the invention to provide compositions of topically active drugsubstances which are effective and convenient. Further objects of theinvention will become clear on the basis of the description of theinvention below, including the examples, and of the patent claims.

SUMMARY OF THE INVENTION

The present invention provides a novel pharmaceutical composition foradministration to the skin or a skin appendage. The compositioncomprises an effective amount of an active ingredient and asemifluorinated alkane (SFA) according to formula RFRH or RFRHRF,wherein RF is a perfluorinated hydrocarbon segment with 20 or lesscarbon atoms and RH is a non-fluorinated hydrocarbon segment with 3 to20 carbon atoms. In a preferred embodiment, the SFA is selected fromF4H5, F4H6, F4H8, F6H6 and F6H8.

It has been surprisingly found by the inventors that SFAs are capable ofdelivering active pharmaceutical ingredients effectively into the deeperlayers of the skin, i.e. into the vital layers below the stratum corneumof the epidermis. Moreover, they are capable of delivering activeingredients deep into nails.

The composition may, for example, be in the form of a solution or anemulsion. They are useful for administering a wide range of topicalagents indicated in a number of cutaneous diseases and conditions.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the amount of tacrolimus found in different layers of theskin (SC=stratum corneum; EP=vital epidermis; DR=dermis; AK=hypodermis)30 min, 300 min, and 1,000 min after administration as an SFA-basedformulation to human skin, relative to the administered dose. Forfurther details, see Example 1.

FIG. 2 shows the amount of tacrolimus found in different layers of theskin (SC=stratum corneum; EP=vital epidermis; DR=dermis; AK=hypodermis)30 min, 300 min, and 1,000 min after administration of a commercialformulation (Protopic® 0.1%) to human skin, relative to the administereddose. For further details, see Example 1.

FIG. 3 shows the penetration profiles of tacrolimus 30 min, 300 min, and1,000 min after administration as an SFA-based formulation to humanskin, respectively. For further details, see Example 1.

FIG. 4 shows the penetration profiles of tacrolimus 30 min, 300 min, and1,000 min after administration a commercial formulation (Protopic® 0.1%)to human skin, respectively. For further details, see Example 1.

FIG. 5 shows the concentration of D,L-α-tocopherol found in differentlayers of the skin (SC=stratum corneum; EP=vital epidermis; DR=dermis)100 min after administration as SFA-based formulation (SFA), Unguentumemulsificans aquosum DAB (WHS), Basiscreme DAC (BC), Lanae alcoholumunguentum aquosum DAB (WWS). For further details, see Example 2.

FIG. 6 shows the concentration of betamethasone-17,21-dipropionate indifferent layers of the skin (SC=stratum corneum; EP=vital epidermis;DR=dermis) 100 min after administration as SFA-based formulation (SFA),Unguentum emulsificans aquosum DAB (WHS), Basiscreme DAC (BC), Lanaealcoholum unguentum aquosum DAB (WWS). For further details, see Example3.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides a novel pharmaceutical composition for use as atopical formulation which is to be administered to the skin or a skinappendage. The composition comprises an effective amount of an activeingredient and a semifluorinated alkane according to formula RFRH orRFRHRF, wherein RF is a perfluorinated hydrocarbon segment with 20 orless carbon atoms and RH is a non-fluorinated hydrocarbon segment with 3to 20 carbon atoms, and in particular, wherein the perfluorinatedsegment is linear and comprises from 4 to 12 carbon atoms, and/or thenon-fluorinated segment is linear and comprises from 4 to 8 carbonatoms.

The invention is based on the surprising discovery that thesemifluorinated alkanes (SFAs) as defined above are highly suitablecarriers for topically administered drugs and capable of delivering suchdrugs very effectively to the layers of the skin below the stratumcorneum, i.e. the viable epidermis, but also to the dermis andhypodermis. This was particularly unexpected in view of Hardung(Doctoral Thesis, Albert-Ludwigs-University of Freiburg, Germany, 2008),who investigated the suitability of SFAs as carriers for topicalmedications using e.g. testosterone as a model drug, and who came to theconclusion that an SFA such as F6H8 (see below for an explanation of theterminology of SFAs) does not affect an improved penetration into theskin, interact with the stratum corneum or exhibit anypenetration-enhancing effect (p. 103, first full paragraph). However, inspite of such discouraging statements in the prior art, the inventorshave further investigated the impact of SFAs and SFA-based carriers onthe delivery of drugs into the deeper layers of the skin and nails usingexcised human skin (in contrast to Hardung who used pig skin), and havenow surprisingly found that SFAs are indeed capable of enhancing thepenetration of drugs into the relevant skin layers, as will be shown inmore detail below.

As used herein, a pharmaceutical composition is any compositioncomprising an active ingredient useful for the diagnosis, prevention,management or therapy of a disease, symptom or health condition and atleast one carrier or excipient. A topical formulation is a compositionwhich is in a form suitable for topical administration. In the presentinvention, the composition is suitable for administration to the skin ora skin appendage of a subject, which subject may be a human or ananimal. The skin may be intact skin or injured, bruised, damaged orotherwise affected skin. A skin appendage is a structure derived fromthe skin, and usually adjacent to it, such as a hair, a sweat orsebaceous gland. Semifluorinated alkanes are linear or branched alkanessome of whose hydrogen atoms have been replaced by fluorine. In apreferred embodiment, the semifluorinated alkanes (SFA's) used in thepresent invention are composed of at least one non-fluorinatedhydrocarbon segment and at least one perfluorinated hydrocarbon segment.Particularly useful are SFA's which have one non-fluorinated hydrocarbonsegment attached to one perfluorinated hydrocarbon segment, according tothe general formula F(CF₂)_(n)(CH₂)_(m)H, or two perfluorinatedhydrocarbon segments separated by one non-fluorinated hydrocarbonsegment, according to the general formula F(CF₂)_(n)(CH₂)_(m)(CF₂)_(o)F.

Another nomenclature which is used herein refers to the above-mentionedSFA's having two or three segments as RFRH and RFRHRF, respectively,wherein RF designates a perfluorated hydrocarbon segment, RH designatesa non-fluorinated segment. Alternatively, the compounds may be referredto as FnHm and FnHmFo, respectively, wherein F means a perfluoratedhydrocarbon segment, H means a non-fluorinated segment, and n, m and ois the number of carbon atoms of the respective segment. For example,F3H3 is used for perfluoropropylpropane. Moreover, this type ofnomenclature is usually used for compounds having linear segments.Therefore, unless otherwise indicated, it should be assumed that F3H3means 1-perfluoropropylpropane, rather than 2-perfluoropropylpropane,1-perfluoroisopropylpropane or 2-perfluoroisopropylpropane.

Preferably, the semifluorinated alkanes according to the generalformulas F(CF₂)_(n)(CH₂)_(m)H and F(CF₂)_(n)(CH₂)_(m)(CF₂)_(o)F havesegment sizes ranging from 3 to 20 carbon atoms, i.e. n, m and o areindependently selected in the range from 3 to 20. SFAs which are usefulin the context of the present invention are also described in EP-A 965334, EP-A 965329 and EP-A 2110126, the disclosure of which documents isincorporated herein.

In a further embodiment, the semifluorinated alkane is a compoundaccording to the formula RFRH, whose segments R_(F) and R_(H) are linearand each—but independently from one another—have from 3 to 20 carbonatoms. In particular, the perfluorinated segment is linear and comprisesfrom 4 to 12 carbon atoms, and/or the non-fluorinated segment is linearand comprises from 4 to 8 carbon atoms. Preferred SFA's include inparticular the compounds F4H5, F4H6, F4H8, F6H4, F6H6, F6H8, and F6H10.Presently most preferred for carrying out the invention are F4H5, F4H6,F6H6 and F6H8.

Optionally, the composition may comprise more than one SFA. It may beuseful to combine SFA's, for example, in order to achieve a particulartarget property such as a certain density, viscosity, or solubilisingcapacity for a particular active ingredient. If a mixture of SFA's isused, it is furthermore preferred that the mixture comprises at leastone of F4H5, F4H6, F4H8, F6H4, F6H6, F6H8, and F6H10, and in particularone of F4H5, F4H6, F4H8, F6H6 and F6H8. In another embodiment, themixture comprises at least two members selected from F4H5, F4H6, F4H8,F6H4, F6H6, F6H8, and F6H10, and in particular at least two membersselected from F4H5, F6H6, F4H8, and F6H8. Furthermore, mixtures of asemifluorinated alkane, e.g. F4H5, with a perfluorinated compound, suchas perfluorooctylbromide or perfluorodecalin, or with another oilymaterial are also contemplated. Other oily materials would, for example,include silicone oils, triglycerides (e.g. native or syntheticmedium-chain triglycerides), or compounds such as isopropyl myristate.In some preferred embodiments, however, perfluorinated compounds areabsent.

Liquid SFA's are chemically and physiologically inert, colourless andstable. Their typical densities range from 1.1 to 1.7 g/cm³, and theirsurface tension may be as low as 19 mN/m. SFA's of the RFRH type areinsoluble in water but also somewhat amphiphilic, with increasinglipophilicity correlating with an increasing size of the non-fluorinatedsegment. Again, for practising the current invention, an SFA having adensity of at least 1.2 g/cm³ should be selected.

Liquid SFA's of the RFRH type are being used commercially for unfoldingand reapplying a retina, for long-term tamponade as vitreous humorsubstitute (H. Meinert et al., European Journal of Ophthalmology, Vol.10(3), pp. 189-197, 2000), and as wash-out solutions for residualsilicon oil after vitreo-retinal surgery. Experimentally, they have alsobeen used as blood substitutes (H. Meinert et al., Biomaterials,Artificial Cells, and Immobilization Biotechnology, Vol. 21(5), pp.583-95, 1993). These applications have established SFA's asphysiologically well tolerated compounds. On the other hand, SFA's havenot been used as excipients in approved drug products as of today.

For allowing convenient administration to the skin or skin appendages,the composition is preferably liquid or semi-solid. As used herein,semi-solid means that a material behaves like a solid upon the exertionof low shear force, but above a certain force threshold, the so-called“yield point”, it behaves like a viscous fluid. Whether liquid orsemi-solid, the composition may represent a single phase (i.e. asolution), or consist of two or more phases.

In a particular embodiment, for example, the composition is in the formof an emulsion. As used herein, an emulsion is a system comprising adispersed (or inner, or emulsified, or discontinuous) liquid phasewithin a continuous (or outer, or coherent) liquid or semisolid phase.The two phases are not miscible. In an O/W-emulsion (also referred to asoil-in-water emulsion), a water-immiscible organic liquid phase, whichdoes not have to be an “oil” by any specific definition, is dispersed ina water-miscible continuous phase which may or may not be substantiallycomprised of water itself. Within emulsions, O/W-emulsions are preferredfor carrying out the invention.

In another embodiment, the composition is in the form of amicroemulsion. A microemulsion is a clear, thermodynamically stable,optically isotropic mixture of a lipophilic component, a hydrophiliccomponent, and an amphiphilic component. Typically, a microemulsionforms spontaneously when the components are combined and mixed with eachother, without requiring high energy input as is normally required forthe formation of an “ordinary” emulsion. Microemulsions may have acolloidal lipophilic phase dispersed in a hydrophilic phase, or ahydrophilic phase colloidally dispersed in a lipophilic phase. The sizeof the dispersed phases is usually in the range from about 5 nm to about400 nm, and most often below about 200 nm. In one of the preferredembodiments of the invention, the particle size is from about 5 nm toabout 100 nm. In terms of its rheological properties, the microemulsionmay be in the form of a liquid or a gel, i.e. in liquid or semisolidform. In a preferred embodiment, the microemulsion is in liquid form.

For microemulsions according to the invention, it is preferred to use anoil phase comprising from about 5 to about 95 wt.-% of SFA, and morepreferably from about 10 to about 80 wt.-% of SFA, the remainder of theoil phase being another oil such as isopropylmyristate. With respect tothe incorporation of aqueous components, surfactants and optionallycosurfactants, commonly known constituents of micro emulsions may beused to formulate the composition.

The composition, whether formulated as a solution, microemulsion orconventional emulsion, may contain any further pharmaceuticallyacceptable ingredients as required or useful in view of the intendedapplication. In a particular embodiment, an additional solvent orco-solvent may be incorporated, for example in order to achieve a highersolubility for a specific active ingredient, to modify the viscosity orstability of the formulation, or to further enhance the cutaneouspenetration of the active ingredient. Such co-solvent should of coursebe selected in type, quality and amount such as to maintain thephysiological tolerability of the formulation. Potentially suitableco-solvents include ethanol, acetone, ethyl acetate, isopropyl alcohol,glycerol, propylene glycol, pentylene glycol, polyethylene glycol,liquid paraffin, triglyceride oils, hydrofluorocarbons such as HFA 134aand/or HFA 227, and liquid mono- or diglycerides. Among these, ethanoland pentylene glycol are among the particularly preferred solvents. Ithas been found that relatively small amounts of ethanol maysubstantially modify the solubilising properties of SFAs and allow theincorporation of a number of active ingredients at higherconcentrations. For example, the composition may comprise up to about 10wt.-% of ethanol, or up to about 5 wt.-% of ethanol. Even at a lowconcentration of up to about 3 wt.-%, ethanol may be a very usefuladditive in the composition, depending on the specific drug substance tobe delivered. Pentylene glycol is also very useful as co-solvent ingeneral and, in particular, it has been found that its incorporation isadvantageous in the formulation of microemulsions.

Moreover, the composition may comprise one or more stabilisers,surfactants (in particular if the composition is in the form of anemulsion or microemulsion), cosurfactants (in particular if it is in theform of a microemulsion), colouring agents, antioxidants (for example,α-tocopherol), thickeners (viscosity-increasing agents such asbentonite), and fragrances.

If a surfactant is present, it may, for example, be selected fromphysiologically acceptable phospholipids (such as phosphatidylcholines);nonionic surfactants such as fatty alcohols, pegylated glycerides,pegylated fatty acids, pegylated fatty alcohols, pegylated sorbitanfatty acid esters, and poloxamers; anionic surfactants such as sodiumlauryl sulphate, docusate sodium, and sodium deoxycholate.

The composition may be prepared by commonly known techniques for themanufacture of pharmaceutical solutions, emulsions, and microemulsions.Depending on their consistency and specific use, they may be presentedin bottles, spray bottles, or tubes.

The active ingredient incorporated within the composition may, inprinciple, be selected from drug substances which are useful for theprevention, management, or therapy of a cutaneous disease or condition.

In one of the specific embodiments, the active ingredient is poorlywater-soluble. In particular, it water solubility is not more than about1 mg/mL. In other preferred embodiments, the water solubility is nothigher than about 0.1 mg/mL, or not more than about 10 μg/mL,respectively. The invention is particularly useful for delivering suchactive ingredients because it allows the administration of effectivedoses in relatively small volumes, which is at least partly due to thesurprisingly high solubilisation capacity of semifluorinated alkanes formany poorly water-soluble drug substances.

For the avoidance of doubt, it is affirmed that the invention is notrestricted to poorly water-soluble drug substances. It has been foundthat certain bioactive agents have substantial aqueous solubility, butcan also be advantageously formulated in SFA-based carriers.

The active ingredient may be selected from drug substances useful forthe prevention and in particular for the management and therapy ofcutaneous diseases and conditions, such as psoriasis, dermatitis,erythema, acne, actinic keratosis, verrucae, and rosacea. The inventionappears generally useful for delivering drug substances of varioustherapeutic categories to the deep layers of the skin, i.e. to the vitalepidermis, the dermis, and/or to the hypodermis which may also beunderstood as being part of the integumentary system. Topicaladministration of the compositions to the skin may result in thedelivery of the drug substance to the stratum corneum, as well aspenetration through the strateum corneum and delivery of the active drugsubstance to these deeper skin layers. Cutaneous diseases or conditionsmay affect the stratum corneum, as well as one or more of the deeperskin layers. The composition may thus be used for the prevention ortreatment of diseases or conditions that affect also the vitalepidermis, the dermis and/or the hypodermis. In one of the specificembodiments, the composition is used in the therapy of psoriasis, andincorporates an active ingredient which may be used to control psoriasisor a symptom thereof. Such active ingredient may, for example, be animmunosuppressant drug, such as a macrolide immunosuppressant. Examplesof potentially suitable macrolide immunosuppressants include tacrolimus,sirolimus, everolimus, tacrolimus, pimecrolimus, ridaforolimus,temsirolimus, zotarolimus, and ciclosporin A; or retinoids such asretinol, retinal, tretinoin, isotretinoin, alitretinoin, etretinate,acitretin, tazarotene, bexarotene and adapalene. Tacrolimus is one ofthe particularly preferred drugs in this group. It has been found by theinventors that tacrolimus can be very effectively (i.e. faster anddeeper than using conventional formulations) delivered to the deep andviable layers of the skin using the composition of the invention (seeExample 1).

As illustrated in Example 1, the composition of the invention may beused to effectively deliver an active ingredient such as a macrolideimmunosuppressant, as for example tacrolimus, to the deeper layers ofthe skin. For example, at least about 5% of the topically administeredamount of drug may be delivered to the vital epidermis within only about30 minutes. The composition may also be used to deliver at least 5% ofthe topically administered drug to the dermis within about 30 minutes,or even at least about 10% of the administered dose. Also in terms ofdelivery to the hypodermis, the composition may be used to deliver atleast 5% of the topically administered amount of drug to this layerwithin the same period of time, or even 10% or more.

Taking into account the typical thickness of the respective skin layers,the composition may be used to deliver within about 30 minutes at leastabout 15% of the topically administered drug dose to a depth of at leastabout 10 μm. In other embodiments, at least about 20% of the dose isdelivered to a depth of at least about 10 μm. The composition may alsobe used to deliver at least about 10% of the topically administered drugto a depth of 100 μm or more within about 30 minutes; or at least about15% or at least about 20% of the dose, respectively.

Another therapeutic indication in which it is of great importance thatthe therapeutic agent penetrates the deeper layers of the affected skinis cutaneous mycosis, such as athlete's foot, candidal vulvovaginitis,and other fungal infections of the integument. in the context, theinvention may be used by preparing and administering an SFA-basedcomposition comprising a suitable topically active antifungal agent, inparticular an azole such as clotrimazole, ketoconazole, or miconazole;or an allylamine, such as terbinafine, naftifine, or butenafine.

In a further embodiment, the disease to be treated is acne, such as acnevulgaris or other forms of acneiform eruptions. In this case, the activeingredient may, for example, be selected from retinoids, such asretinol, retinal, tretinoin, isotretinoin, alitretinoin, etretinate,acitretin, tazarotene, bexarotene and adapalene; antiinfective agentssuch as azithromycin, erythromycin, clindamycin, or minocycline; orspironolactone.

According to another embodiment, the invention is used in the treatmentof actinic keratosis, which is a premalignant condition of the skincharacterised by the presence of thick, scaly, or crusty patches. Inthis case, the active ingredient may be selected from NSAIDs such asdiclofenac, ketoprofene and flurbiporofene, but also 5-fluorouracil andimiquimod.

Moreover, the invention is suitable for the treatment of viralinfections of the skin, such as herpes simplex, herpes zoster, andvaricella. For this indication, an antiviral compound is incorporated inthe SFA-based composition, such as acyclovir, penciclovir, or docosanol.

In a further embodiment, the composition of the invention comprises alocal anaesthetic agent such as benzocaine, butamben, dibucaine,lidocaine, oxybuprocaine, pramoxine, proparacaine, proxymetacaine, andtetracaine. Such composition may be used to relieve pain or itchingcaused by conditions such as sunburn or other minor burns, insect bitesor stings, or in preparation of minor surgical procedures.

A large number of cutabeous diseases and conditions involve symptomsrelated to inflammatory responses, such as most forms of dermatitis,including atopic dermatitis, seborrhoeic dermatitis, allergicdermatitis, contact dermatis and the like, but also certain forms ofpruritus, psoriasis, allergies, nummular eczema, xerotic eczema, lichensclerosis, lichen planus, discoid lupus, chapped feet, lichen simplexchronicus, poison ivy reactions, alopecia areata, nummular eczema etc.Often these conditions require therapy or symptomatic control withtopical corticosteroids, which can also be formulated and administeredadvantageously according to the invention. Suitable corticosteroid whichmay be incorporated in an SFA-based composition include amcinonide,betamethasone dipropionate, clobetasol propionate, desonide,desoximetasone, diflorasone diacetate, fluocinolone acetonide,fluocinonide, flurandrenolide, fluticasone propionate, halcinonide,halobetasol proprionate, hydrocortisone, hydrocortisone butyrate,hydrocortisone valerate, mometasone furoate, prednicarbate, andtriamcinolone acetonide.

The invention is further illustrated by the following examples.

EXAMPLES Example 1

Tacrolimus was dissolved in a liquid carrier comprising F6H8 and 3.8wt.-% ethanol to obtain a clear solution having a tacrolimusconcentration of 1 mg/mL.

Excised human skin was obtained from a breast reduction surgical centre.Skin samples were washed with isotonic sodium chloride solution. Thesubcutaneous adipose layers were removed. Circular pieces of 20 mm indiameter (3.14 cm²) were punched out, supported by filter gauze, andmounted in FRANZ diffusion cells in such a way that the dermal side wasin contact with the acceptor fluid (PBS pH 7.4), which was continuouslystirred and kept at 32° C. Loss through evaporation was avoided by meansof a glass lid.

On each skin sample (epidermal side), an amount of 20 μL of thetacrolimus solution was placed and spread out evenly. After a testingtime of 30 min, 300 min, or 1,000 min, respectively, the remainingtacrolimus solution on the samples was collected and the skin sampleswere removed. From each specimen, three small punch biopsies of 6 mmdiameter (0.2827 cm²) were obtained and frozen. At −40° C., the punchbiopsies were cryomicrotomised as follows:

-   -   1 slice of 10 μm thickness, corresponding to the stratum corneum    -   4 slices of 20 μm each, corresponding to the vital epidermis    -   5×5 slices of 40 μm each, corresponding to 5 layers of dermis

All slices and the remaining stumps were extracted, and the amount oftacrolimus was determined by HPLC-MS. Moreover, the acceptor fluid andthe fluid removed from the stratum corneum after each test were alsoanalysed for tacrolimus. Each test was performed in triplicate, usingskin from three different donors.

For comparison, a similar series of experiments using a commercialtacrolimus ointment (Protopic® 0.1%, Astellas Pharma, 20 mg per skinsample and FRANZ cell) was conducted.

In result, it was found that the penetration of tacrolimus into therelevant skin layers was clearly superior in the case of the solutionaccording to the invention. For example, already after 30 minutesapprox. 9% of the administered dose was found in the vital epidermis,and approx. 12% was found in the layers of the dermis (see FIG. 1). Forcomparison, the commercial formulation delivered only about 2.5% of thedose to the vital epidermis and only about 5% to the dermis during thesame period of time (see FIG. 2). After 1,000 minutes, the amount in thedermis was about 30% of the dose in the case of the solution of theinvention (see FIG. 1), but only about 12.5% in the case of thecommercial preparation (see FIG. 2). At all times, the penetration depthwas much higher for the composition of the invention (see FIG. 3) thanfor the commercial product (see FIG. 4).

Example 2

A solution of radiolabeled D,L-α-tocopherol in F6H8 having aconcentration of 10 mg/mL was prepared and used for a series of skinpenetration experiments in analogy to Example 1, using excised humanskin, with a few differences to the procedure of Example 1 as describedbelow.

The amount of the formulation applied to each skin sample was 6 mg. Thetest period was 100 min. After the testing period, the stratum corneumof each sample was removed by the film stripping method (20 strips).Slices of the vital epidermis and the dermis were however obtained bymicrotomy at −40° C.

For comparison, three pharmacopoeial ointment formulations incorporatingthe same concentration of D,L-α-tocopherol were prepared and tested:“Unguentum emulsificans aquosum” (DAB), “Basiscreme” (DAC), and “Lanaealcoholum unguentum aquosum” (DAB).

In result, it was surprising to find that the SFA-based formulationresulted in a much higher concentration of tocopherol in the vitalepidermis than the three conventional formulations (see FIG. 5).

Example 3

The series of experiments of Example 2 were repeated in analogy, butusing radiolabeled betamethasone-17,21-dipropionate as active ingredientat a concentration of 1 mg/mL instead of tocopherol. Another differencewas that the SFA-based formulation of the drug contained 2.5 wt.-%ethanol.

Here it was found that the drug concentration in the skin layers washighest in the case of the SFA-based formulation of the invention, witha clear difference between the tested formulations with respect to thestratum corneum and the vital epidermis, but somewhat lessdifferentiation in the dermis (see FIG. 6).

Example 4

An SFA-based microemulsion with tacrolimus was prepared. Basically, 10mg of tacrolimus were dissolved in 1.5 mg of Tween® 80 and 2 g of Brij®30. Then 2 g of 1,2-pentandiole were added, followed by 0.3 g of F6H8and 0.7 g of isopropyl myristate. After gentle stirring, 4 g of waterwere added and the mixture was homogenised until it had a transparentappearance.

The invention claimed is:
 1. A method for treating a disease orcondition of the skin comprising topically delivering an activeingredient to layers of the skin below the stratum corneum selected fromthe vital epidermis, the dermis and/or the hypodermis by topicallyadministering to the skin a topical pharmaceutical compositionconsisting of an effective amount of an active ingredient and asemifluorinated alkane according to formulaRFRH wherein RF is a linear perfluorinated hydrocarbon segment with 4 to12 carbon atoms, and RH is a linear alkyl group with 4 to 8 carbonatoms, and optionally one or more surfactants, co-surfactants,stabilizers, coloring agents, anti-oxidants, thickeners, fragrances,and/or an additional semifluorinated alkane, a perfluorinated compound,a silicone oil, isopropyl myristate, or a physiologically acceptableco-solvent selected from ethanol, acetone, ethyl acetate, isopropylalcohol, glycerol, propylene glycol, pentylene glycol, polyethyleneglycol, liquid paraffin, triglyceride oils, hydrofluorocarbons, andliquid mono- or diglycerides; wherein the active ingredient is selectedfrom an immunosuppressant agent, a retinoid, an antifungal agent, ananti-infective agent, an antiviral agent, a local anesthetic, anon-steroidal anti-inflammatory drug (NSAID), a corticosteroid,5-fluorouracil and imiquimod; wherein the disease or condition of theskin is selected from psoriasis, dermatitis, erythema, eczema, pruritus,lichen sclerosis, lichen planus, discoid lupus, chapped feet, lichensimplex chronicus, poison ivy, alopecia areata, allergy, acne, actinickeratosis, fungal infections, bacterial infections, viral infections,verrucae, rosacea, ichthyosis, callus, sunburn, minor burns, insectbites and insect stings.
 2. The method of claim 1, wherein thesemifluorinated alkane is selected from F4H5, F4H6, F4H8, F6H6 and F6H8,wherein F means a perfluorinated hydrocarbon segment, H means anon-fluorinated segment, and the number is the number of carbon atoms ofthe respective segment.
 3. The method of claim 1, wherein thecomposition is in the form of a liquid or semisolid solution, emulsion,or microemulsion.
 4. The method of claim 1, wherein the co-solvent isselected from ethanol, acetone, ethyl acetate, isopropyl alcohol,glycerol, propylene glycol, pentylene glycol, polyethylene glycol,liquid paraffin, triglyceride oils, hydrofluorocarbons, and liquid mono-or diglycerides.
 5. The method of claim 1, wherein the disease orcondition of the skin is selected from psoriasis, dermatitis, erythema,eczema, allergy, acne, actinic keratosis, fungal infections, bacterialinfections, viral infections, verrucae, or rosacea.
 6. The method ofclaim 1, wherein the disease or condition of the skin is by psoriasis,and wherein the active ingredient is selected from immunosuppressants orretinoids.
 7. The method of claim 6, wherein the immunosuppressant isselected from tacrolimus, sirolimus, everolimus, tacrolimus,pimecrolimus, ridaforolimus, temsirolimus, zotarolimus, and ciclosporinA.
 8. The method of claim 6, wherein the retinoid is selected fromretinol, retinal, tretinoin, isotretinoin, alitretinoin, etretinate,acitretin, tazarotene, bexarotene and adapalene.
 9. The method of claim1, wherein the disease or condition of the skin is acne, and wherein theactive ingredient is selected from retinoids or anti-infective agents.10. The method of claim 9, wherein the retinoid is selected fromretinol, retinal, tretinoin, isotretinoin, alitretinoin, etretinate,acitretin, tazarotene, bexarotene and adapalene.
 11. The method of claim9, wherein the anti-infective agent is selected from azithromycin,erythromycin, clindamycin, or minocycline.
 12. The method of claim 1,wherein the disease or condition of the skin is actinic keratosis, andwherein the active ingredient is selected from diclofenac,5-fluorouracil and imiquimod.
 13. The method of claim 1, wherein thedisease or condition of the skin is a cutaneous disease involvingsymptoms related to inflammatory responses and the active ingredient isa corticosteroid.
 14. The method of claim 1, wherein the disease orcondition of the skin is pain or itching and the active ingredient is alocal anesthetic agent.
 15. The method of claim 1, wherein the diseaseor condition of the skin is a fungal infection and the active ingredientis an anti-fungal agent.
 16. The method of claim 1, wherein the diseaseor condition of the skin is a viral infection, and the active ingredientis an anti-viral agent.
 17. The method of claim 1, wherein i. at leastabout 5% of the topically administered amount of active ingredient isdelivered to the vital epidermis within about 30 minutes, or ii. atleast about 5% of the topically administered amount of active ingredientis delivered to the dermis within about 30 minutes; or iii. at leastabout 10% of the topically administered amount of active ingredient isdelivered to the dermis within about 30 minutes; or iv. at least about5% of the topically administered active ingredient is delivered to thehypodermis within about 30 minutes; or v. at least about 10% of thetopically administered active ingredient is delivered to the hypodermiswithin about 30 minutes.
 18. The method of claim 1, wherein at leastabout 15% of the topically administered active ingredient is deliveredto a depth of at least 10 μm within about 30 minutes.
 19. The method ofclaim 1, wherein at least about 20% of the topically administered activeingredient is delivered to a depth of at least 10 μm within about 30minutes.
 20. The method of claim 1, wherein at least about 10% or atleast 15% or at least about 20% of the topically administered activeingredient is delivered to a depth of at least 100 μm within about 30minutes.
 21. The method of claim 1, wherein the topical pharmaceuticalcomposition is a solution.
 22. The method of claim 1, wherein the activeingredient is a drug substance useful for the prevention, management ortherapy of a cutaneous disease or disorder.
 23. The method of claim 22,wherein the cutaneous disease or disorder is selected from psoriasis,dermatitis, erythema, acne, actinic keratosis, verrucae, rosacea,eczema, allergy, fungal infections and viral infections.